Automated immunoanalyzer system for performing diagnostic assays for autoimmune and infectious diseases

Abstract

A quantitative method for diagnosing an autoimmune disease or an infectious disease comprising performing an automated diagnostic assay, comprising: incubating a capture reagent with a streptavidin-coated medium to form a solid phase complex, wherein the capture reagent is a biotinylated autoantigen or infectious disease antigen; washing the solid phase complex to remove excess capture reagent; incubating the solid phase complex with a serum sample to form an immune complex; washing the immune complex to remove any unbound sample; incubating the immune complex with a conjugate to create an immune-conjugate complex; washing the immune-conjugate complex to remove any unbound conjugate; introducing a substrate capable of generating a quantifiable response; and calibrating the response generated from introducing the substrate.

What is claimed is: 
     
         1 . A method of performing a diagnostic assay, comprising:
 a) combining a capture reagent-coated separation medium, wherein the separation medium comprises fluorescently-labelled particles, with an analyte-containing sample in a reaction cuvette;   b) binding luminescent labels to the separation media in proportion to a number of bound analyte molecules, to form a complex;   c) measuring an initial fluorescent signal associated with an initial quantity of the fluorescently-labelled particles;   d) washing the complex;   e) adding a luminescing reagent or substrate to the cuvette to generate a luminescent signal in proportion to the number of bound analyte molecules,   f) measuring the luminescent signal and a final fluorescent signal associated with a final quantity of the fluorescently-labelled particles in the cuvette;   g) calculating a ratio of the final fluorescent signal to the initial fluorescent signal to obtain a bead retention ratio; and   h) adjusting a calibrated quantifiable response for bead retention by adjusting the luminescent signal by the bead retention ratio to calculate a reported value.   
     
     
         2 . The method of  claim 1 , wherein the fluorescently-labelled particles are magnetic. 
     
     
         3 . The method of  claim 1 , wherein the analyte-containing sample is a serum sample. 
     
     
         4 . The method of  claim 1 , wherein the analyte-containing sample is a plasma sample. 
     
     
         5 . The method of  claim 1 , wherein the capture reagent will be bound by analyte molecules in the analyte-containing sample. 
     
     
         6 . The method of  claim 6 , wherein the analyte molecules are capture reagent-specific immunoglobulins. 
     
     
         7 . The method of  claim 1 , wherein the binding of luminescent labels is mediated by a labeled anti-immunoglobulin conjugate. 
     
     
         8 . The method of  claim 7 , wherein the anti-immunoglobulin conjugate comprises horseradish peroxidase (HRP), Alkaline Phosphatase (ALP), or beta galactosidase (BGAL). 
     
     
         9 . The method of  claim 1 , wherein the fluorescently-labelled particles have been coated with streptavidin. 
     
     
         10 . The method of  claim 9 , wherein the capture reagent has been biotinylated and bound to the fluorescently-labelled particles through a biotin-streptavidin interaction. 
     
     
         11 . The method of  claim 6 , wherein the immunoglobulins are analyte-specific human immunoglobulin G (IgG). 
     
     
         12 . The method of  claim 6 , wherein the immunoglobulins are analyte-specific human immunoglobulin M (IgM). 
     
     
         13 . The method of  claim 6 , wherein the immunoglobulins are analyte-specific human immunoglobulin A (IgA) 
     
     
         14 . The method of  claim 6 , wherein the immunoglobulins are analyte-specific human immunoglobulin E (IgE). 
     
     
         15 . The method of  claim 1 , wherein binding luminescent labels to the separation media comprises incubating a labeled anti-immunoglobulin conjugate with the capture reagent-coated separation medium. 
     
     
         16 . The method of  claim 15 , wherein the conjugate is in a diluent including polyethylene glycol. 
     
     
         17 . The method of  claim 1 , wherein measuring the fluorescent signals and the luminescent signal takes place within an optics box. 
     
     
         18 . The method of  claim 1 , further comprising entering the fluorescence and luminescence measurements into an algorithm to generate a bead retention adjusted relative light unit signal. 
     
     
         19 . The method of  claim 18 , further comprising comparing the generated bead retention adjusted relative light unit signal to a calibration curve relative light unit signal.